Author:
Date:
2020-10-20
[Abstract] Ascorbic acid (AsA) and gluthathione (GSH) are two key components of the antioxidant machinery of eukaryotic and prokaryotic cells. The cyanobacterium Synechocystis sp. PCC 6803 presents both compounds in different concentrations (AsA, 20-100 μM and GSH, 2-5 mM). Therefore, it is important to have precise and sensitive methods to determine the redox status in the cell and to detect variations in this antioxidants. In this protocol, we describe an improved method to estimate the content of both antioxidants (in their reduced and oxidized forms) from the same sample obtained from liquid cultures of Synechocystis sp. PCC 6803.
[摘要] [摘要] 抗坏血酸(AsA)和明胶硫(GSH)是真核细胞和原核细胞抗氧化机制的两个重要组成部分。蓝藻Synechocystis sp.pcc6803以不同浓度(AsA,20-100μM和GSH,2-5mm)呈现这两种化合物。因此,有精确和敏感的方法来确定细胞中的氧化还原状态和检测这种抗氧化剂的变化是很重要的。在该方案中,我们描述了一种改进的方法,用以从Synechocystis sp.pcc6803的液体培养中获得的同一样品中估算两种抗氧化剂(以还原形式和氧化形式)的含量。 [背景] 细胞中的氧化还原状态可以被多种因素改变,产生氧化应激。我们使用该方案来量化暴露于50℃高温下的Synechocystis sp. PCC 6803中的GSH和AsA含量。如拟南芥所述,热胁迫导致抗氧化剂含量下降,并通过铁作用引起细胞死亡(Distefano et al., 2017;Aguilera等人,2019年预印本)。虽然本方案是针对Synechocystis sp. PCC 6803制定的,但也可用于测定其他蓝藻中GSH和AsA的含量。蓝藻中AsA含量很低,正常条件下uM含量在一定范围内,某些处理下pM含量在一定范围内。因此,我们提出了这个敏感的方法与一个改进的细胞裂解程序。
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